Optogenetic control of endothelial cells

Abstract

The invention features methods for regulating vascular properties by controlling the membrane properties of endothelial cells using optogenetics and light. The invention features methods to transport therapeutics across the vascular barrier into tissues such as the brain and the lung, with high spatial and temporal precision, and for controlling vascular properties such as vascular tone, arterial diameter, and vascular growth.

What is claimed is: 
     
         1 . A method for changing the permeability of endothelial cells, said method comprising the step of contacting endothelial cells expressing optogenetic reagents with light, wherein said light activates said optogenetic reagents and thereby changes the permeability of said endothelial cells. 
     
     
         2 . A method for changing the permeability of endothelial cells, said method comprising: a) infecting endothelial cells with recombinant viruses comprising a recombinant nucleic acid encoding said optogenetic reagent to produce infected endothelial cells; and b) contacting said infected endothelial cells with light, wherein said light activates said optogenetic reagents and thereby changes permeability of said endothelial cells. 
     
     
         3 . (canceled) 
     
     
         4 . The method of  claim 2 , wherein said optogenetic reagents are selected from the group consisting of ChR1, ChR2, VChR1, ChR2 C128A, ChR2 C128S, ChR2 C128T, ChD, ChEF, ChF, ChIEF, NpHR, eNpHR, Arch 3.0, Arch T 3.0, Mac 3.0, melanopsin, chimeras of these proteins and natural and engineered variants thereof. 
     
     
         5 - 6 . (canceled) 
     
     
         7 . The method of  claim 2 , wherein said recombinant nucleic acid is encapsidated within a recombinant virus selected from the group consisting of recombinant adeno-associated virus (AAV), recombinant retrovirus, recombinant lentivirus, recombinant poxvirus, recombinant rabies virus, recombinant pseudo-rabies virus, and recombinant herpes simplex virus, and human immunodeficiency virus (HIV). 
     
     
         8 . The method of  claim 2 , wherein said recombinant nucleic acid encoding said optogenetic reagents further encodes a fiduciary marker, that when expressed identifies cells infected by the recombinant virus. 
     
     
         9 . The method of  claim 8 , wherein said fiduciary marker is a fluorescent protein. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 2 , wherein said light may be applied using a laser or a light emitting diode, and wherein application of light may be restricted to a defined spatial region of the body. 
     
     
         12 . The method of  claim 11 , wherein said light is delivered using a fiber optic cable or catheter. 
     
     
         13 - 15 . (canceled) 
     
     
         16 . The method of  claim 2 , wherein said change in permeability results in a closing of the blood-brain barrier. 
     
     
         17 . The method of  claim 16 , wherein said closing of blood-brain barrier decreases delivery of elements from the endothelial cells to the brain. 
     
     
         18 . The method of  claim 2 , wherein the light is shined in a specific region of the brain for a specific period of time, providing spatial and temporal control of the opening of the blood-brain barrier. 
     
     
         19 . The method of  claim 2 , wherein said method is used to regulate vascular tone in a subject. 
     
     
         20 . The method of  claim 2 , wherein said method is used to regulate arterial diameter. 
     
     
         21 - 22 . (canceled) 
     
     
         23 . The method of  claim 2 , wherein said method is used to regulate vascular growth. 
     
     
         24 - 29 . (canceled) 
     
     
         30 . The method of  claim 2 , wherein said endothelial cells are part of a blood-air barrier in the lung. 
     
     
         31 . The method of  claim 30 , wherein said method is used to treat a disease of the lung. 
     
     
         32 . The method of  claim 31 , wherein said disease is a lung cancer. 
     
     
         33 . (canceled)